Urothelial cell response to bacterial infection

尿路上皮细胞对细菌感染的反应

• 批准号: 6732071
• 负责人: ROBERT Evan HURST
• 金额: $ 14.65万
• 依托单位: UNIVERSITY OF OKLAHOMA HLTH SCIENCES CTR
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-05-01 至 2006-10-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6732071
• 关键词: Enterococcus; antibiotics; apoptosis; bacteria infection mechanism; bacterial genetics; biological signal transduction; cell adhesion; cell differentiation; drug resistance; functional /structural genomics; gene expression; inflammation; microarray technology; polymerase chain reaction; terminal nick end labeling; tissue /cell culture; urinary bladder epithelium; urinary tract infection; urology

DESCRIPTION (provided by applicant): The urothelium maintains an active set of defenses against bacterial infection. These include static defenses in the form of surface molecules that inhibit bacterial attachment and active responses to eliminate an established bacterial infection. The long-term aim of this study is to elucidate the mechanisms by which antibacterial defenses are modulated by infection. Our hypothesis is that in an environment of increasing bacterial resistance to antibiotics, understanding the natural mechanisms of defense should permit their augmentation as a means to prevent or treat infection, particularly in vulnerable populations. Enterococcus spp. rank second among the leading cause of bacterial UTIs. The high antibiotic resistance prevalent among this genus makes treatment of enterococcal infections a therapeutic challenge. Previous research in our laboratory has established models for urothelium grown in 3 dimensions that closely mimics both the morphology and functional genomics of the bladder urothelium. In this model system, the environment of the urothelial cells can be controlled precisely and specific modulatory proteins can be added, or the actions of specific genes can be inhibited or augmented in a realistic cell culture environment. We will ask two broad questions: 1. How does the urothelium respond to bacterial infection? 2. Can altering expression of specific genes in the urothelium modulate the response to bacterial infection? This approach will integrate closely with independent bacterial and animal model studies carried out by other members of this collaborative research group. Our model permits mechanistic investigations in which the role of specific molecules in the response of urothelium to infection can be tested. Our main objective for this period of support is to expand on current cancer-related cDNA array technology in our laboratory to investigate genome-wide changes in the expression of human urothelial cells in 3-dimensional culture as they experience infection with enterococcus. We plan to use the Clontech system on plastic consisting of over 8,000 named human genes. The aims are to cluster genes according to their behavior over time, identify the key signaling and response pathways that are involved in the response of urothelial cells to infection, and learn how to manipulate and interpret large-scale genomic data. These data will form an essential element in either an R01 or a program project and will permit the PI to develop a new line of urologic research.

描述（由申请人提供）：尿路上皮维持一套活跃的防御细菌感染。这包括表面分子的静态防御，以抑制细菌附着和主动反应的形式消除已建立的细菌感染。本研究的长期目标是阐明感染调节抗菌防御的机制。我们的假设是，在细菌对抗生素耐药性增加的环境中，了解防御的自然机制应该允许它们作为预防或治疗感染的一种手段，特别是在脆弱人群中。肠球菌在引起细菌性尿路感染的主要原因中排名第二。该属中普遍存在的高抗生素耐药性使肠球菌感染的治疗成为一项治疗挑战。我们实验室之前的研究已经建立了三维生长的尿路上皮模型，该模型密切模仿膀胱尿路上皮的形态和功能基因组学。在这个模型系统中，可以精确地控制尿路上皮细胞的环境，添加特定的调节蛋白，或者在真实的细胞培养环境中抑制或增强特定基因的作用。我们将提出两个广泛的问题：1。尿路上皮对细菌感染有何反应？2. 改变尿路上皮中特定基因的表达能否调节对细菌感染的反应？该方法将与该合作研究小组其他成员进行的独立细菌和动物模型研究紧密结合。我们的模型允许进行机制研究，其中可以测试特定分子在尿路上皮对感染的反应中的作用。我们在此期间的主要目标是扩展我们实验室中目前与癌症相关的cDNA阵列技术，以研究人类尿路上皮细胞在三维培养中受到肠球菌感染时表达的全基因组变化。我们计划在含有8000多个已命名人类基因的塑料上使用克隆技术系统。目的是根据基因随时间的行为进行聚类，确定尿路上皮细胞对感染的反应中涉及的关键信号和反应途径，并学习如何操纵和解释大规模基因组数据。这些数据将构成R01或计划项目的基本要素，并将允许PI开发泌尿学研究的新路线。